Images

ab16053 staining GAP43 in murine dorsal root ganglia by Immunocytochemistry/ Immunofluorescence.Cells were fixed with methanol, permeabilized using 0.1% saponin/PBS, blocked with 4% BSA for 30 minutes at 25°C and then incubated with ab16053 at a 1/500 dilution for 14 hours at 4°C. The secondary antibody used was a DyLight 488 conjugated goat anti-rabbit polyclonal used at a 1/100 dilution.

Immunofluorescent staining obtained with ab16053 Rabbit polyclonal to GAP43 in rat dorsal root ganglion, 2 weeks after a spinal nerve injury. The staining is localised in the cytoplasm of some lesioned neurons (arrow in B) and in many axons (arrow heads in B). Image B (X40 objective) is a higher magnification of A (X20). The staining disappeared completely when the antibody was pre-incubated with the immunising (blocking) peptide (ab16379).

Protocol details: Rats were intracardially perfused with 4% paraformaldehyde, DRG tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. IHC free floating was performed on the fixed, frozen and cut (20µm sections) rat tissue. Primary antibody ab16053 was incubated overnight at toom temperature at 1/300. Secondary antibody Alexa fluor 488 was incubated at 1/1000 for 2 hours at room temperature.

We would recommend using ab16053 in place of the discontinued item. Ab16053 has been tested in and is guaranteed in IF on mouse samples. This product has also been featured in 5 publications which we are aware of and has excellent Abreviews. More infor...

Your protocol looks good. The one recommendation I could make would be to increase the dilution of the primary and secondary (especially) to see if that improves the staining. You can try the primary at 1:250 a...

Thank you for calling us and for alerting us to the problem you are experiencing with anti-GAP43 antibody (ab16053). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.